کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1936528 | 1050693 | 2008 | 6 صفحه PDF | دانلود رایگان |
Immobilization of divalent Nickel cations provides a tool for affinity purification of proteins containing hexahistidine tags. During experiments to generate single-stranded DNA aptamers to immobilized proteins we inadvertently identified DNA sequences with affinity for Nickel–nitrilotriacetic acid (Ni2+–NTA) magnetic beads. Analysis of these aptamers revealed that affinity for the Ni2+–NTA support requires only single-stranded sequences with multiple adenosine residues. Bound nucleic acids can be eluted with imidazole. A single-stranded dA20 affinity tag (but not other homopolymer sequences) is sufficient for immobilization of double-stranded DNA PCR products on Ni2+–NTA magnetic beads. Addition of an rA20 sequence to an RNA transcript allowed its affinity capture on Ni2+–NTA magnetic beads, suggesting an approach for purification of poly(A) mRNA.
Journal: Biochemical and Biophysical Research Communications - Volume 366, Issue 2, 8 February 2008, Pages 420–425